TFAM is an autophagy receptor that limits inflammation by binding to cytoplasmic mitochondrial DNA.

When cells are stressed, DNA from energy-producing mitochondria can leak out and drive inflammatory immune responses if not cleared. Cells employ a quality control system called autophagy to specifically degrade damaged components. We discovered that mitochondrial transcription factor A (TFAM)-a protein that binds mitochondrial DNA (mtDNA)-helps to eliminate leaked mtDNA by interacting with the autophagy protein LC3 through an autolysosomal pathway (we term this nucleoid-phagy). TFAM contains a molecular zip code called the LC3 interacting region (LIR) motif that enables this binding. Although mutating TFAM's LIR motif did not affect its normal mitochondrial functions, more mtDNA accumulated in the cell cytoplasm, activating inflammatory signalling pathways. Thus, TFAM mediates autophagic removal of leaked mtDNA to restrict inflammation. Identifying this mechanism advances understanding of how cells exploit autophagy machinery to selectively target and degrade inflammatory mtDNA. These findings could inform research on diseases involving mitochondrial damage and inflammation.

Detection of ribophagy in yeast and mammals.

Ribophagy, the cellular process wherein ribosomes are selectively self-digested through autophagy, plays a pivotal role in maintaining ribosome turnover. Understanding the molecular regulatory mechanisms governing ribophagy is pivotal to uncover its significance. Consequently, the establishment of methods for detecting ribophagy becomes important. In this protocol, we have optimized, enriched, and advanced existing ribophagy detection techniques, including immunoblotting, fluorescence microscopy, and transmission electron microscopy (TEM), to precisely monitor and quantify ribophagic events. Particularly noteworthy is the introduction of TEM technology for yeast ribophagy detection. In summary, the delineated methods are applicable for detecting ribophagy in both yeast and mammals, laying a solid foundation for further exploring the physiological importance of ribophagy and its potential implications in diverse cellular environments.

Association between time of onset and Omicron BA.2 RNA clearance time in children: a retrospective cohort study.

Background: The association between the time of onset [time from the date of detection of a positive real-time reverse-transcription polymerase chain reaction (RT-PCR) to the date of detection of a positive RT-PCR in the first child] and viral RNA clearance time (time from first positive RT-PCR to two consecutive negative RT-PCR) remains unclear. Our study aimed to evaluate their association. That can provide a reference for the number of nucleic acid tests. Methods: We conducted a retrospective analysis of children diagnosed with Omicron BA.2 infection at Fujian Medical University Affiliated First Quanzhou Hospital between March 14, 2022 (date the first child in the outbreak was found positive for RT-PCR) and April 9, 2022 (date the last child was found positive for RT-PCR). We used the electronic medical record to extract demographic data, symptoms, radiology and laboratory findings, treatments, and viral RNA clearance time. The 282 children were divided equally into 3 groups according to the time of onset. We calculated the factors affecting viral RNA clearance time by univariate and multivariate analysis. We used the generalized additive model to investigate the relationship between the time of onset and viral RNA clearance time. Results: 46.45% of children were female. Fever (62.06%) and cough (15.60%) were the dominant onset symptoms. We found no serious cases and all children were cured. The median time to viral RNA clearance was 14 days (IQR 12-17 days), with a range of 5 to 35 days. After adjustment for potential confounders, the viral RNA clearance time was reduced by 2.45 (95% CI: 0.85, 4.04) days in the 7-10 days group and by 4.62 (95% CI: 2.38, 6.14) days in > 10 days group compared to the ≤6 days group. There was a non-linear association between the time of onset and viral RNA clearance time. Conclusions: Time of onset was non-linearly associated with Omicron BA.2 RNA clearance time. During the first 10 days of the outbreak, viral RNA clearance time decreased with increasing onset date. After 10 days of the outbreak, viral RNA clearance time did not decrease with increasing onset date.

Robotic guidance for percutaneous placement of triangular osteosynthesis in vertically unstable sacrum fractures: a single-center retrospective study.

BACKGROUND: To evaluate the effectiveness and safety of robot-aided percutaneous triangular osteosynthesis combined with close reduction for vertically unstable sacrum fractures (VUSFs). METHODS: The data on 21 patients of the VUSF were retrospectively analyzed from November 2016 to January 2021. According to Denis classification, there were 3 cases in zone I, 11 cases in zone II, and 7 case in zone III. The main perioperative indicators were recorded. The maximal angulation and displacement deviations of the screws were analyzed by comparing the planned trajectory with the actual position. Postoperative X-ray radiographs and CT scans were obtained for evaluating the reduction quality. Functional outcome was scored with Majeed criterion. RESULTS: Fourteen patients of the unilateral VUSF and 7 patients of the bilateral VUSF underwent unilateral and bilateral triangular osteosynthesis with robotic assistance, respectively. No intraoperative neurovascular injuries and postoperative infection occurred. All patients were followed up for at least 12 months. The average operation time of posterior pelvic ring was 111.4 min, with the mean intraoperative bleeding of 110.5 ml. A total of 58 pedicle and iliosacral screws were implanted with robotic assistance. Of those, 52 screws were in the cancellous bone except 4 pedicle and 2 iliosacral screws cutting the cortical bone. The angulation and displacement deviations of the screws were 4.2° ± 2.5° and 1.7 ± 0.9 mm, respectively. The average displacement of the sacral fracture was reduced from 19.7 mm preoperatively to 3.1 mm postoperatively. According to Matta's criterion, the reduction quality was graded as "excellent" in 13 patients and "good" in 8. All sacral fractures healed within 6 months except one fracture with nonunion. The mean Majeed score at the last follow-up was 89.6. CONCLUSIONS: Robot-aided triangular osteosynthesis combined with close reduction provide a safe and reliable option for percutaneous treatment of the fresh VUSF, with a high accuracy of iliosacral and pedicle screw implantation except insertion of iliac screws. Meanwhile, the technique may help to reduce incision-related complications.

Comparative retrospective study of triangular osteosynthesis with and without robotic assistance for unilateral unstable sacral fractures combined with lumbosacral junction injuries.

BACKGROUND: To compare the clinical efficacy of unilateral unstable sacral fractures (USFs) involving the lumbosacral region treated with and without robot-aided triangular osteosynthesis (TOS). METHODS: Patients of the unilateral USF combined with the ipsilateral lumbosacral junction injury (LSJI) treated with TOS were retrospectively analyzed and divided into two groups: the robot group (TOS with robotic assistance) and the conventional group (TOS with open procedure). Screw placement was assessed using the modified Gras criterion. Patients were followed up with routine visits for clinical and radiographic examinations. At the final follow-up, clinical outcomes were recorded and scored using the Majeed scoring system. RESULTS: Eleven patients in the robot group and seventeen patients in the conventional group were recruited into this study. Significant differences in surgical bleeding (P < 0.001) and fluoroscopy time (P = 0.002) were noted between the two groups. Operation time (P = 0.027) and fracture healing time (P = 0.041) was shorter in the robot group. There was no difference in postoperative residual displacement between the two groups (P = 0.971). According to the modified Gras criterion, the percentages of grade I for sacroiliac screws in the two groups were 90.9% (10/11) and 70.6% (12/17), and for pedicle screws were 100% (11/11) and 100% (17/17), respectively. The rate of incision-related complications was 0% (0/11) in the robot group and 11.8% (2/17) in the conventional group. Statistical differences were shown on the Majeed criterion (P = 0.039), with higher scores in the robot group. CONCLUSION: TOS with robotic assistance for the treatment of unilateral USFs combined with ipsilateral LSJIs is safe and feasible, with the advantages of less radiation exposure and fewer incision-related complications.

The Kocher-Langenbeck approach combined with robot-aided percutaneous anterior column screw fixation for transverse-oriented acetabular fractures: a retrospective study.

OBJECTIVE: Transverse-oriented acetabular fractures (TOAFs), including transverse, transverse with posterior wall and T-shaped fractures, are always challenging for double-column reduction and fixation with minimally invasive method. The purpose of this study is to compare the therapeutic effects of robot-aided percutaneous anterior column screw fixation versus minimally invasive anterior plate fixation for TOAFs based on the Kocher-Langenbeck (K-L) approach. METHODS: Patients suffering TOAFs that were fixed by robot-aided percutaneous anterior column screw fixation or minimally invasive anterior plate fixation associated with posterior fixation via the K-L approach were divided into two groups: group A (screw fixation) and group B (plate fixation). Surgical time, blood loss, incision length for anterior fixation and complications were recorded. Fracture reduction quality was evaluated using criteria described by Matta. Fracture healing was assessed on the series of pelvic radiographs at each follow-up. Functional outcomes were investigated using the modified Postel Merle D'Aubigne score at the final follow-up. RESULTS: Twenty-nine patients with TOAFs, including 12 patients in group A and 17 patients in group B, were evaluated for study eligibility. The mean surgical time of anterior fracture fixation was 18.7 ± 4.6 min in group A and 33.4 ± 5.0 min in group B (P < 0.001). The amount of intraoperative blood loss was 615.6 ± 178.7 ml in group A and 719.3 ± 199.0 ml in group B (P < 0.001). Incision length for anterior fixation was 9.0 ± 1.8 mm in group A and 81.2 ± 7.3 mm in group B (P < 0.001). The complications related to the surgery of anterior column only occurred in group B (lateral femoral cutaneous nerve palsy in 1 patient and groin discomfort in 1 patient). No significant differences in reduction quality, hospital stay, fracture healing time and functional results were noted between the two groups. CONCLUSION: The K-L approach combined with robot-aided anterior column screw fixation is a safe and effective option for TOAFs. Compared with minimally invasive anterior plate fixation, robot-aided screw fixation has obvious advantages on surgical time, blood loss, and invasiveness. The K-L approach combined with minimally invasive anterior plate fixation can also be a reliable alternative for TOAFs, with the similar reduction quality and functional results.

Emergence of novel avian origin H7N9 viruses after introduction of H7-Re3 and rLN79 vaccine strains to China.

In early 2021, roughly 6 months after the H7N9 H7-Re3 and H7N9 rLN79 vaccine strains were introduced into China, we monitored a number of H7N9 subtype avian influenza viruses, which could have escaped vaccine-induced immunity in live poultry markets (LPMs) in Yunnan, Hebei, Shanxi and Guangdong provinces, China. To investigate whether these viruses were a novel H7N9 variant of highly pathogenic avian influenza (HPAI) virus and whether they had the potential for further spread, we characterized the genetic evolution, antigenic divergence and pathogenicity of the viruses in the context of vaccine immunity. The results show further diversification in the HA gene of newly isolated HPAI H7N9 viruses compared with antigenic variants that emerged after the period of 2017-2019. There were clear antigenic differences between current vaccines and these viruses, and SPF broilers under vaccine protection could not resist virus challenges. Our study demonstrates that the current vaccine has insufficient protective capacity against the novel H7N9 variants under experimental conditions. A novel H7N9 immune escape virus has emerged. Faced with potential outbreaks, we should strengthen surveillance and update vaccine strains.

Comparison of Outcomes Following TiRobot-Assisted Sacroiliac Screw Fixation with Bone Grafting and Traditional Screw Fixation without Bone Grafting for Unstable Osteoporotic Sacral Fracture: A Single-Center Retrospective Study of 33 Patients.

BACKGROUND This retrospective study from a single center aimed to compare patient outcomes following TiRobot-assisted sacroiliac screw fixation and bone grafting with traditional screw fixation without bone grafting in 33 patients with unstable osteoporotic sacral fracture (UOSF). MATERIAL AND METHODS Patients with UOSF were included and divided into 2 groups: a TiRobot-assisted surgical group with 18 patients (robot-aided sacroiliac screw fixation and bone grafting) and a standard surgical group with 15 patients (free-hand screw fixation without bone grafting). T values of bone mineral density (BMD) £-2.5 standard deviation (SD) were diagnosed as osteoporosis. Screw positioning and fracture healing time were evaluated. Functional outcomes were investigated at the final follow-up. RESULTS There were no statistically significant differences in screw positioning; however, there were satisfactory positioning rates in 94.4% (17/18) of patients in the TiRobot-assisted surgical group and 73.3% (11/15) in the standard surgical group. The advantages with TiRobot on surgical time of screw placement, fluoroscopy frequency, and total drilling times were noted (P=0.000). The nonunion rates were 5.6% (1/18) in the TiRobot-assisted surgical group and 33.3% (5/15) in the standard group (P=0.039). Healing time in the union cases had a significant difference (P=0.031). Functional outcome scores in the TiRobot-assisted surgical group were superior to that in the standard group (P=0.014). CONCLUSIONS The findings showed that TiRobot-assisted sacroiliac screw fixation and bone grafting was a safe and effective surgical treatment option that had a reduced radiation dose and improved fracture healing, when compared with standard screw fixation without bone grafting.

Robot-Aided Minimally Invasive Lumbopelvic Fixation in Treatment of Traumatic Spinopelvic Dissociation.

OBJECTIVE: To investigate the surgical strategy, safety, and efficacy of close reduction and robot-aided minimally invasive lumbopelvic fixation in treatment of traumatic spinopelvic dissociation. METHODS: Data of 32 patients (21 males and 11 females) with traumatic spinopelvic dissociation treated by lumbopelvic fixation with robot-aided minimally invasive technique or conventional open procedure in our institution from March 2010 to April 2019 were retrospectively analyzed, and divided into robot group and control group. Intraoperative blood loss, surgical time, fluoroscopy frequency, total drilling times, infection rate, hospitalization time, and sacral fracture healing time were reviewed. Radiographs and computed tomography (CT) scans were totally acquired to evaluate the reduction quality, residual fracture displacement, and Gras classification on screws insertion after surgery. According to the Majeed scoring system, functional outcome was assessed for each patient at the final follow-up. RESULTS: There were 12 patients in the robot group and 20 patients in the control group with no significant difference about the demographic data. The average surgical time was 148.3 ± 40.5 min with intraoperative blood loss of 142.5±36.7 mL in the robot group and 185.0 ± 47.8 min with 612.5 ± 182.7 mL in the control group (P = 0.034, P = 0.000). The robot group had a shorter mean hospitalization time at 19.9 ± 7.0 days compared to the control group with 28.6 ± 5.4 days (P = 0.010). The fluoroscopy frequency was 35.4 ± 3.0 in the robot group and 45.5 ± 3.6 in the control group (P = 0.000) and total drilling times were 7.1 ± 1.1 and 9.6 ± 1.3 (P = 0.000), respectively. The infection rate was 0% (0/12) in the robot group and 15% (3/20) in the control group (P = 0.159). According to the Gras classification on screw positioning, there were 11 cases in Grade I and 1 case in Grade II in the robot group, and 14 cases in Grade I and 6 cases in Grade II in the control group. All the patients were followed up consecutively for at least 12 months, with an average follow-up period of 17.1 ± 3.6 months. All sacral fractures healed with an average time of 3.8 ± 0.6 months in the robot group and 4.7 ± 0.7 months in the control group (P = 0.000). According to Majeed functional assessment investigation, the mean score of the patients was 87.2 ± 4.0 in the robot group and 83.1 ± 4.5 in the control group (P = 0.015). CONCLUSIONS: Robot-aided minimally invasive lumbopelvic fixation for traumatic spinopelvic dissociation is a safe and feasible option with advantages of less intraoperative blood loss, less radiation damage, less hospitalization time, and better functional outcome.

Clinical research on minimally invasive internal fixation for the treatment of anterior ring injury in tile C pelvic fracture.

The aim of this study is to explore the clinical outcome and indications in treating anterior ring injury of Tile C pelvic fracture with minimally invasive internal fixation.We retrospectively reviewed 18 patients (aged 25-62, 34.2 ±â€Š7.4) with 26 pelvic anterior ring injuries of Tile C pelvic fracture treated with minimally invasive internal fixation in our hospital were from January 2012 to August 2016. Two cases were pubic symphysis diastasis, 15 were anterior ring fracture (7 were bilateral), and 1 was vertical displacement of pubic symphysis associated with pubic ramus fracture. According to Tile classification, 8, 4, and 6 cases were types C1, C2, and C3, respectively. All patients accepted the operation of pelvic fractures on both rings, while the anterior ring injuries were treated with minimally invasive internal fixation. The period from injury to operation was 5 to 32 days (11.2 ±â€Š3.7). Four patients had pubic symphysis diastasis or pelvic anterior ring fracture medial obturator foramen reduced with modified Pfannenstiel incision and fixed with cannulated screws, 14 patients (22 fractures) had a fractured lateral obturator foramen reduced with modified Pfannenstiel incision associated with small iliac crest incision and fixed with locking reconstruction plates. Clinical data, such as operation time, intraoperative bleeding, Matta standard to assess the reduction quality of fracture, and complications, were collected and analyzed.The operation time ranged from 30 to 65 minutes (42.8 ±â€Š18.7), and the intraoperative bleeding volume was 30 to 150 mL (66.5 ±â€Š22.8). All cases were continuously followed-up for 16 to 42 months (30.2 ±â€Š4.6). All fractures were healed between 3 and 9 months postoperatively (4.9 ±â€Š2.7 months). According to the Matta standard assessment, 18, 7, and 1 cases were excellent, good, and fair, respectively, with a 96.2% (25/26) rate of satisfaction. Neither reduction loss, fixation failure, nor infection occurred; complications included 1 patient with fatal liquefaction, 1 patient had lateral femoral cutaneous nerve injury, and 1 patient complained of discomfort in the inguinal area due to fixation stimulation.Minimally invasive internal fixation for pelvic anterior ring injury in Tile C pelvic fracture has the advantages of less damage, safer manipulation, less complications, and good prognosis.

Reference markers of bone turnover for prediction of fracture: a meta-analysis.

OBJECTIVE: To explore whether bone turnover biomarkers (BTMs), i.e., C-terminal telopeptide of type I collagen (CTX) and procollagen type I aminoterminal propeptide (PINP), are associated with fracture. METHODS: We searched electronic database including PubMed, Embase and Cochrane Library, and the reference lists of relevant articles published from inception to August 22, 2018. An updated meta-analysis was performed to assess the prediction value of CTX and PINP in fracture. RESULTS: Nine articles met our inclusion criteria and were included in the meta-analysis. The crude and adjusted effect size between PINP and fracture were extracted from two and five studies, respectively. PINP was not associated with fracture incidence without adjusting covariates (crude GR, 1.03; 95% CI, 0.91-1.17). After adjusting for potential confounders, PINP demonstrated a significant positive association with fracture (adjusted GR, 1.28; 95% CI, 1.15-1.42). In the subgroup analysis of studies after adjusting covariates, there were significant associations in women. Both the crude (1.16, 95%CI, 1.04-1.20) and adjusted GR (1.20, 95%CI, 1.05-1.37) shown positive relationships between CTX and fracture, which were extracted from four and six studies, separately. The sensitivity analysis confirmed the stability of the results. In the subgroup analysis of studies after adjusting covariates, there were significant associations in the subgroups of elderly, female, and hip fracture patients. CONCLUSIONS: Our results indicate a statistically significant but modest association between BTMs (s-PINP or s-CTX) and future fracture risk after adjusting for BMD and clinical risk factors. The causal relationship between the two clinical conditions requires future validation with more standardized studies. REGISTRATION NUMBER: CRD42018107879.

Analysis of Crisis Management of Medical Disputes in China and Australia: A Narrative Review Article.

At present, the increasing trend of medical disputes has become a serious problem in the work of medical institutions, and directly affects the social stability and orderly development of the institutions. We searched the literature on medical disputes and crisis management in China and Australia within PubMed (2010-2019), China Knowledge Network (CNKI, 2010-2019), and Wanfang Data Knowledge Service Platform (2010-2019). There are several drawbacks in the management of hospital medical disputes in China: 1) the knowledge of crisis is unilateral and not systematic; 2) there are too much stereotype thoughts in crisis management; 3) the crisis attribution is too simple; the crisis impact assessment is insufficient. It is worth learning from Australia's system, including the legal system, relevant non-governmental organizations, and doctor-patient communication. In view of the malpractice existing in China's medical dispute management, Australia's legal system for handling disputes, doctor-patient communication and other aspects are worthy of our reference. In particular, the construction of third-party supervision and mediation institutions and the prevention of doctor-patient disputes should be optimized.

Polyelectrolyte Complex Nanoparticles Based on Methoxy Poly(Ethylene Glycol)-B-Poly (ε-Caprolactone) Carboxylates and Chitosan for Delivery of Tolbutamide.

In this study, a novel chitosan (CS)-modified nanoparticles (NPs) were developed to orally deliver tolbutamide (TOL). Methoxy poly(ethylene glycol)- b-poly(ε-caprolactone) carboxylates (mPEG2000-b-PCL4000) was synthesized via an esterification reaction. CS-modified mPEG2000-b-PCL4000-COOH NPs (CS@NPs) were fabricated by injecting mPEG2000-b-PCL4000-COOH NPs suspension (1.0 mg/mL) into CS solution (1.0 mg/mL, pH 5.0). Fourier transform infrared spectroscopy (FTIR) spectra were used to confirm the obtaining of mPEG2000-b-PCL4000-COOH. Transmission electron microscope (TEM) was carried out to observe morphology of all NPs. Nano ZS90 Malvern ParticleSizer were used to monitor the size distribution of obtained NPs. Thermogravimetry analysis (TGA) was performed to investigate the thermostability of CS@NPs. In vitro TOL release profiles were carried out in pH 1.2 and 7.4 buffers. FTIR spectra confirmed the obtaining of mPEG2000-b-PCL4000-COOH. TGA curves indicated that the protection of CS shells improved the thermostability of mPEG2000-b-PCL4000-COOH NPs. Cell tests indicated the CS@NPs had no obvious cytotoxicity, and they were easily taken up by 293T cells. In vitro release profiles showed that 91.0 ± 1.9% of encapsulated TOL were released from TOL-CS@NPs in pH 7.4 buffer. Therefore, the positive potential of CS@NPs could increase their combining capacity with intestinal mucosal cells. Finally, these NPs would improve the bioavailability of hydrophobic drugs.

Internal Fixation of Complicated Acetabular Fractures Directed by Preoperative Surgery with 3D Printing Models.

The purpose of this article is to evaluate the efficacy and feasibility of preoperative surgery with 3D printing-assisted internal fixation of complicated acetabular fractures. A retrospective case review was performed for the above surgical procedure. A 23-year-old man was confirmed by radiological examination to have fractures of multiple ribs, with hemopneumothorax and communicated fractures of the left acetabulum. According to the Letounel and Judet classification, T-shaped fracture involving posterior wall was diagnosed. A 3D printing pelvic model was established using CT digital imaging and communications in medicine (DICOM) data preoperatively, with which surgical procedures were simulated in preoperative surgery to confirm the sequence of the reduction and fixation as well as the position and length of the implants. Open reduction with internal fixation (ORIF) of the acetabular fracture using modified ilioinguinal and Kocher-Langenbeck approaches was performed 25 days after injury. Plates that had been pre-bent in the preoperative surgery were positioned and screws were tightened in the directions determined in the preoperative planning following satisfactory reduction. The duration of the operation was 170 min and blood loss was 900 mL. Postoperative X-rays showed that anatomical reduction of the acetabulum was achieved and the hip joint was congruous. The position and length of the implants were not different when compared with those in preoperative surgery on 3D printing models. We believe that preoperative surgery using 3D printing models is beneficial for confirming the reduction and fixation sequence, determining the reduction quality, shortening the operative time, minimizing preoperative difficulties, and predicting the prognosis for complicated fractures of acetabulam.

Low­molecular weight fucoidan inhibits the differentiation of osteoclasts and reduces osteoporosis in ovariectomized rats.

Fucoidan is a type of sulfated polysaccharide isolated from seaweed. The present study used ovariectomized Sprague­Dawley rats, which were treated with fucoidan. The effects of fucoidan on bone metabolism, density and microarchitecture were assessed using micro­computed tomography (CT), histomorphometric analysis, biochemical markers of bone metabolism (Serum procollagen type I N propeptide and C­terminal telopeptide­1) and tests of mechanical competence of the femur. In addition, the effects of low­molecular weight fucoidan (LMWF) on in vitro cultured osteoclasts were examined, in order to determine the mechanisms underlying LMWF­induced osteoclastic inhibition. In ovariectomized rats, LMWF increased femoral bone density. Micro­CT scan also revealed that LMWF prevented microarchitectural deterioration and histomorphometric analysis determined that LMWF increased trabecular bone number and reduced the surface of bone resorption. In addition, LMWF reduced the high bone turnover rate, and improved the mechanical properties of the femur in ovariectomized rats. In vitro experiments revealed that LMWF inhibited the receptor activator of nuclear factor κB ligand (RANKL) and macrophage colony­stimulating factor­induced differentiation of RAW264.7 cells into tartrate­resistant acid phosphatase (TRAP)­positive osteoclasts, and reduced the bone resorption surface of the osteoclasts. Reverse transcription­quantitative polymerase chain reaction demonstrated that LMWF inhibited mRNA expression of TRAP, matrix metallopeptidase­9, nuclear activator of activated T­cells 1, and osteoclast­associated immunoglobulin­like receptor, which are components of the signaling pathway for osteoclast differentiation. LMWF had no effect on RANK mRNA expression. In conclusion, the present study confirmed that LMWF inhibited osteoclast differentiation and bone resorption, and may be a potential treatment for osteoporosis in ovariectomized rats.

The anthraquinone derivative Emodin inhibits angiogenesis and metastasis through downregulating Runx2 activity in breast cancer.

Emodin (EMD) is an anthraquinone derivative extracted from the root and rhizome of Rheum palmatum L. which exhibits a range of activities, including anti-bacterial, antitumor, diuretic and vasorelaxant effects. The ability to inhibit metastasis and angiogenesis was shown in previous pharmacological studies, but clear information to address EMD affecting angiogenesis and metastasis in human breast cancer is still lacking. In the present study, we evaluated a possible role for EMD in angiogenesis and metastasis induced by breast cancer cells. It was revealed here that EMD attenuated tumor cell-induced metastasis and angiogenesis both in vitro and in vivo. Furthermore, it was found that these inhibitory effects were caused by MMPs and VEGFR-2 inhibition in metastatic breast cancer cells and endothelial cells, respectively. Western blot analysis showed reduction of Runx2 activation in the EMD-treated cells. ELISA based Runx2 transcription factor assay showed that the interaction between Runx2 and target sequences was inhibited by EMD. Our findings suggested that the inhibitory effects of EMD on tumor-induced metastasis and angiogenesis were caused by MMPs and VEGFR-2 inhibition, which may be associated with the downregulation of Runx2 transcriptional activity.

Codonolactone, a sesquiterpene lactone isolated from Chloranthus henryi Hemsl, inhibits breast cancer cell invasion, migration and metastasis by downregulating the transcriptional activity of Runx2.

Metastasis is the most insidious aspect of breast cancer, but effective strategies to control this malignant process are still lacking. In previous studies, we screened over 200 extracts from plants of genus Chloranthaceae by bioactivity-guided fractionation, and found that Codonolactone (CLT) exhibited potential antimetastatic properties in breast cancer cells. This sesquiterpene lactone was isolated from Chloranthus henryi Hemsl, and is also found in other medical herbs, such as Codonopsis pilosula, Atractylodes macrocephala Koidz and others. Here, we report that CLT inhibited the ability of invasion and migration in metastatic breast cancer cells. Furthermore, CLT exhibited significant suppression on formation of lung metastatic foci of breast cancer in vivo. We next investigated the mechanism of CLT-induced metastasis inhibitory effects in breast cancer cells. A significant inhibition on activity and expression of MMP-9 and MMP-13 was observed. Moreover, data from western blotting, Runx2 transcription factor assay and chromatin immunoprecipitation assay showed that binding ability of Runx2 to sequences of the mmp-13 promoter was inhibited by CLT. Collectively, these findings suggested that the antimetastatic properties of CLT in breast cancer were due to the inhibition of MMPs, which might be associated with a downregulation of Runx2 transcriptional activity.

The dineolignan from Saururus chinensis, manassantin B, inhibits tumor-induced angiogenesis via downregulation of matrix metalloproteinases 9 in human endothelial cells.

Manassantin B (MB) is a neolignan isolated from Saururus chinensis that exhibits a range of activities, including anti-inflammatory, antiseptic and antitumor activity. MB was recently found to affect cell adhesion and expression of several adhesion molecules. Based on the important roles of these adhesion molecules in angiogenesis, we evaluated a possible role for MB in tumor-induced angiogenesis in endothelial cells (ECs). In the present study, we found that MB blocked tumor-induced tube formation of ECs and significantly inhibited the invasion of ECs through the reconstituted basement membrane. MB suppressed the activity of matrix metalloproteinases (MMPs) and downregulated the expression of matrix metalloproteinases 9. Western blotting showed reduction of RUNX2 activation by MB. RUNX2 transcription factor assay and chromatin immunoprecipitation assay showed that the interaction between RUNX2 and target sequences in the matrix metalloproteinases 9 promoters was inhibited by MB. Our findings suggested that the inhibitory effects of MB on tumor-induced angiogenesis were caused by matrix metalloproteinases 9 inhibition, which was associated with the downregulation of RUNX2 transcriptional activity.

[Clinical characteristics and surgical management of Eyres type V coracoid fracture combined with superior shoulder suspensory complex injury].

OBJECTIVE: To investigate the clinical characteristics of Eyres type V coracoid fracture combined with superior shoulder suspensory complex (SSSC) injuries, and the effectiveness of open reduction and fixation. METHODS: Between March 2004 and July 2012, 13 patients with Eyres type V coracoid fracture and SSSC injuries were treated. There were 10 males and 3 females with an average age of 41 years (range, 23-59 years). Injury was caused by falling from height in 4 cases, by traffic accident in 6 cases, and by impact of the heavy weight in 3 cases. The interval from injury to operation was 3-10 days (mean, 5.2 days). SSSC injuries included 9 cases of acromioclavicular joint dislocation, 5 cases of clavicular fractures, and 4 cases of acromion fractures. The coracoid fractures were fixed with cannulated screws; the acromioclavicular joint dislocations were fixed with hook plate (6 cases) or Kirschner wires (2 case) except 1 untreated case; the clavicular fractures were fixed with anatomical locking plate (3 cases) and hook plate (2 cases); the acromion fractures were fixed with cannulated screws (1 case), Kirschner wires (2 cases), or both of them (1 case). RESULTS: The mean operation time was 158.0 minutes (range, 100-270 minutes), and the mean intraoperative blood loss was 207.7 mL (range, 150-300 mL). The other patients obtained primary healing of incision except 1 patient who had inflammation around incision, which was cured after change dressing. All patients were followed up for 22.6 months on average (range, 17-35 months). All fractures achieved union at a mean time of 3.6 months (range, 2-6 months). No nerve injury and implant fixation failure complications were observed. At last follow-up, the Constant score and the disability of the arm, shoulder, and hand (DASH) score had a significant improvement when compared with scores at pre-operation (P < 0.05). The shoulder range of motion in flexion, abduction, and external rotation at last follow-up were significantly higher than those at pre-operation (P < 0.05). CONCLUSION: Eyres type V coracoid fracture associated with SSSC injuries usually results in the instability of the shoulder. With individual surgical treatment, the satisfactory function and good effectiveness can be obtained.

[Detection of protein secretion and expression and osteogenic activity of adipose tissue-derived stem cells after in vitro transfection of vascular endothelial growth factor].

OBJECTIVE: To examine vascular endothelial growth factor (VEGF) protein secretion and expression and explore the osteogenic activity of adipose tissue-derived stem cells (ADSCs) after transfection of human VEGF. METHODS: The ADSCs were isolated from human adipose tissue after the digestion of collagenase.After identification by flow cytometry, the cells were cultured and passaged in nutritive medium. Gene sequence encoding human VEGF mature peptide was obtained by Trizol reagent method from human vascular tissue. Target gene VEGF was connected with bicistronic expression vector containing green fluorescent protein to form pSELECT-GFP zeo-VEGF for transfecting 2nd, 3rd, 4th, 5th generation ADSCs mediated by liposome. The transfection results were verified under fluorescence microscope. VEGF protein secretion by transfected cells was detected by enzyme-linked immunosorbent assay (ELISA). Second-generation transfected ADSCs were cultured under osteogenic conditions.The supernatant levels of alkaline phosphatase (ALP) and osteocalcin (OC) were detected. RESULTS: Liposome-mediated VEGF target gene fragment could transfect ADSCs successfully. ELISA quantitative detection showed that VEGF mRNA expression levels in supernatant of the transfected group was significantly higher than the control group. And there were significant differences. After osteogenic culturing, the detections of ELISA, real-time PCR and Western blot showed that the secretion of ALP and OC of VEGF transfected group was significantly higher than that of empty vector transfected and blank cell groups.And there were significant differences (P < 0.01). CONCLUSION: After transfected by liposome-mediated VEGF target gene fragment, human ADSCs can express biologically active VEGF mRNA in vitro continuously and effectively. Directional differentiation capacity of transfected ADSCs is significantly enhanced.